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Detection of genetically modified rice: a construct-specific real-time PCR method based on DNA sequences from transgenic Bt rice
Authors:Dietrich Mäde  Christine Degner  Lutz Grohmann
Affiliation:(1) Landesamt für Verbraucherschutz Sachsen-Anhalt, Dezernat 33, Freiimfelder Str. 66/68, 06112 Halle, Germany;(2) Bundesamt für Verbraucherschutz und Lebensmittelsicherheit, Abteilung Gentechnik, Referat 405, Mauerstr. 39-42, 10117 Berlin, Germany
Abstract:Genetically modified rice varieties developed in China are close to approval for agricultural cultivation and production. However, so far no method has been reported for specific detection of transgenic varieties of this crop. In the present study, rice seeds assumed to consist of field-tested Bt rice (‘Anti-pest Shanyou 63’ and ‘Anti-pest Jinyou 63’) were used as reference material to determine transgenic DNA sequences. The transition between the cryIA(b) and cryIA(c) fusion gene and the nopaline synthase terminator (nos) sequence was used to develop a construct-specific real-time PCR based detection method. This Bt rice specific detection system was combined with a recently published quantitative real-time PCR method for the rice-specific (Oryza sativa L.) reference gene gos9. The complete PCR assay for detection of transgenic Bt rice was in-house validated and the limit of quantification was found to be below 0.1% Bt rice relative to the rice content. Application of the PCR assay should allow more precise detection of transgenic rice varieties in imported food products which are so far not approved in the EU.
Keywords:Genetically modified organism  Rice (Oryza sativa L  )  Bt toxin gene  Reference gene  Real-time PCR  Detection method
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